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Benjamin Gully

Thesis

Evaluation of cavitands and nanoparticles as additives for protein crystallisation

Summary

This thesis involves the structural analysis of proteins via X-ray diffraction, a technique employed to gain atomic resolution detail of a proteins structure. Which in turn aids understanding of how a protein mediates the processes it is involved in.

A rate-limiting step in this technique is the generation of protein crystals (an ordered arrangement of protein molecules akin to how sucrose molecules are arranged in sugar crystals), which can take years. My thesis is in the experimental use of nano-scale additives applied to protein crystallisation in an attempt to crystallise difficult proteins and solve their structures.

If you see figure 1. with a lego brick representing the protein, I can explain. We aim to improve the process either by causing the lego bricks together hoping some slot together forming the foundation of a crystal (nucleation based approach). Or alternatively I can use additives to make a section more stable and thus more likely to build up into a crystal (cavitand based approach).

Why my research is important

This research is important because protein structures are so important, we can learn so much on from a protein structure but the current attrition from selection of a protein to crystallising it is ~98%. The potential of nano-scale materials applied as outlined here is vast, any improvement in solubility, stability and potential crystallisability of proteins would aid the structural analysis of proteins that are currently difficult or impossible to crystallise.

For example one such protein we are working on binds to RNA in an ordered, repeating fashion. Initial crystalisation attempts have failed but nano-additives are proving useful, if we can crystallise this protein we will be able to understand how this protein binds the RNA. This knowledge could afford a designer protein that we can use to bind strategic RNA targets, if successful this would have huge biotechnological applications.

Funding

Australian Postgraduate Award (Industry) funded by the Australian Research Council and Chimere Pearls.

Scholarship for International Research Fees funded by the University of Western Australia.